Compounds and methods for treatment of primary biliary cholangitis

ABSTRACT

The present invention relates to, inter alia, methods of treatment and combinations of (R)-2-(7-(4- cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1) useful for the treatment of primary biliary cholangitis (PBC). In some embodiments, the methods further comprise administering Compound 1, or a pharmaceutically salt, solvate, or hydrate thereof, in combination with a compound selected from the group consisting of: an antihistamine (diphenhydramine), cholestyramine (questran, prevalite), rifampin, an opioid antagonist (naloxone), pilocarpine (isopto carpine, salagen), cevimeline (evox-ac), calcium and/or vitamin D supplement, and vitamin A, D, E and/or K supplement. Other embodiments, relate to titration packages for enabling compliance with a regimen of changing dosage of a medication over a period of time for the treatment of primary biliary cholangitis (PBC).

FIELD OF THE INVENTION

The present invention relates to, inter alia, methods of treatment and combinations of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1) useful for the treatment of primary biliary cholangitis (PBC). In some embodiments, the methods further comprise administering Compound 1, or a pharmaceutically salt, solvate, or hydrate thereof, in combination with a compound selected from the group consisting of: an antihistamine (diphenhydramine), cholestyramine (questran, prevalite), rifampin, an opioid antagonist (naloxone), pilocarpine (isopto carpine, salagen), cevimeline (evoxac), calcium and/or vitamin D supplement, and vitamin A, D, E and/or K supplement. Other embodiments relate to titration packages for enabling compliance with a regimen of changing dosage of a medication over a period of time for the treatment of primary biliary cholangitis (PBC).

BACKGROUND OF THE INVENTION

The sphingosine-1-phosphate (S1P) receptors 1-5 constitute a family of G protein-coupled receptors with a seven-transmembrane domain. These receptors, referred to as S1P₁ to S1P₅ (formerly termed endothelial differentiation gene (EDG) receptor-1, -5, -3, -6, and -8, respectively; Chun et al., Pharmacological Reviews, 54:265-269, 2002), are activated via binding by sphingosine-1-phosphate, which is produced by the sphingosine kinase-catalyzed phosphorylation of sphingosine. S1P₁, S1P₄, and S1P₅ receptors activate Gi but not Gq, whereas S1P₂ and S1P₃ receptors activate both Gi and Gq. The S1P₃ receptor, but not the S1P₁ receptor, responds to an agonist with an increase in intracellular calcium.

The compound (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1) is a potent (EC₅₀ cAMP, 0.093 nM (human)) and selective (EC₅₀ β-arrestin, 6.10 nM (S1P₁), >10,000 nM (S1P₂), >10,000 nM, (S1P₃), 147 nM (S1P₄), and 24.4 nM (S1P₅)), orally available investigational drug candidate for the S1P₁ receptor.

In preclinical studies, Compound 1 showed calculated lymphocyte lowering IC₅₀ values in four different species: 0.101 µM (mouse), 0.051 µM (rat), 0.058 µM (dog), and 0.098 µM (monkey). Notably, the calculated lymphocyte lowering IC₅₀ values reflect total plasma concentration wherein Compound 1 is highly protein bound (97.8% human, 98.0% rat). Compound 1 was shown to be efficacious in the murine experimental autoimmune encephalomyelitis (EAE) model that mimics multiple sclerosis. Prophylactically, Compound 1 prevented the onset and severity of disease relative to vehicle up to day 25, at which time dosing was discontinued. All treatment arms went on to develop severe disease. Therapeutic administration of Compound 1 was also examined. Treatment began at day 18, by which time all animals had developed severe disease. Compound 1 was administered from day 18 to day 37 and showed to reverse the disease relative to vehicle and was similar to the efficacy observed with fingolimod (i.e., GILENYA® was approved in September 2010 for the treatment of individuals with relapsing forms of multiple sclerosis). Similarly, Compound 1 was efficacious in a collagen induced arthritis (CIA) model. Prophylactic oral administration in female Lewis rats resulted in a significant reduction in ankle diameters on day 17 following a daily oral dose and was similar to that observed in rats treated with fingolimod or methotrexate. Improvement in histological parameters in the knees and ankles of CIA rats was also observed, suggesting that inhibiting lymphocyte entry into arthritic joints with Compound 1 treatment suppresses CIA in rodents. Additional details can be found in the following, PCT application, serial number PCT/US2009/004265, filed 22 Jul. 2009 (International Publication Number WO2010/011316); PCT application, serial number PCT/US2011/000153, filed 27 Jan. 2011 (International Publication Number WO2011/094008); and Buzard: D. J., et al., ACS Med. Chem. Lett. 2014, 5, 1313—1317; each hereby incorporated by reference in its entirety.

S1P is a signaling sphingolipid required by lymphocytes to exit the lymphoid tissue and enter the bloodstream via a chemotactic gradient. The S1P1 receptor is a physiological mediator which has been shown to regulate lymphocyte recirculation between lymphoid tissue and blood. Binding and internalization of the S1P1 receptor may result in lymphocyte retention within lymphoid tissue, with subsequent reduction in peripheral lymphocyte count and lymphocyte availability for recruitment to sites of inflammation. S1P1 receptor surface expression is required for S1P gradient-mediated lymphocyte migration out of lymphoid tissue into the circulation (Brinkmann V., Nat Rev Drug Discov 2010 November; 9(11):883-97).

Primary biliary cholangitis is a chronic cholestatic liver disease of unknown cause. Progressive bile-duct injury from portal and periportal inflammation may result in progressive fibrosis and eventual cirrhosis. Immunohistochemical staining of T lymphocytes in portal and periportal areas in PBC patients shows CD4-positive and CD8-positive T cells (T-cell populations known to be modulated by S1P1 interaction). In addition to T cells, natural killer cells also appear to play a role in PBC. Resting as well as activated NK cells express S1P1, S1P4, and S1P5 receptors. The S1P5 receptor has been reported to be involved in chemotaxis of NK cells (Jenne et al 2009). Allende et al. demonstrated a role for S1P4 in neutrophil trafficking in that S1P lyase deficient mice (Sgp11-/- GrS1pr1) did not significantly differ from the single mutant Sgp11-/-mice in blood levels of lymphocytes and neutrophils and in serum concentrations of pro-inflammatory cytokines). In contrast, mice that lacked both S1P lyase and S1P4 (Sgp11-/- S1pr4-/-) had significantly lower blood neutrophil counts and serum pro-inflammatory cytokines than the single-mutant Sgpll-/-mice (Allende et al. 2011). These data suggest involvement of the S1P4 receptor in neutrophil trafficking, which may be relevant given reports suggesting a role for IL-33 in enhancing the migration of neutrophils in the progression PBC (Sun et al. 2014). Avoidance of S1P2 interaction is important due to the reported links between S1P2 modulation in cholangiocarcinoma (CCA), possibly driven by conjugated bile acid modulation of the S1P2 receptor on cholangiocytes (Liu et al 2014). Inhibition of S1P1, but not S1P2 receptors, has been shown to reduce bile salt (glycochenodeoxycholic acid)-induced apoptosis in rat hepatocytes, indicating potential therapeutic benefits in PBC (Karimian et al., Biochim Biophys Acta. 2013 Dec;1832(12):1922-9). Further, it has been reported that S1P1 may be involved in processes promoting liver fibrosis, suggesting that blockade of the S1P1 pathway may help attenuate liver fibrosis (Yang L et al., J Hepatol. 2013 Jul;59(1):114-23). Modulation of the S1P1, S1P4, and S1P5 receptors therefore represents a target profile for treating PBC.

However, many of the S1P modulators that are currently on the market or in clinical development have reportedly shown evidence of elevating liver transaminases upon chronic administration. For example, liver enzyme elevation has been seen for GILENYA (fingolimod), siponimod, ponesimod, GSK2018682, and ozanimod (Gergely et al., British J of Pharm 2012; 167:1035-1047; D’Ambrosio et al., Therapeutic Advances in Chronic Disease 2016; 7(1):18-33; Cohen et al., 32^(nd) Congress of ECTRIMS 2016 Sep 14-17; Xu et al., Am College of Clinical Pharm 2014; 3(3): 170-178). In fact, the product label for GILENYA (fingolimod) contain warnings and precautions about hepatic effects. Further, there is pathophysiological evidence of a reduction in drug metabolizing enzyme activities in patients with PBC, along with general liver impairment effects (as evidenced by elevated liver transaminases) (Reshetnyak, World J of Gastroenterology 2015 July 7; 21(25):7683-7708). As such, the safety of a S1P modulator in a patient population with impaired liver function is unknown.

Described herein is a proof-of-concept clinical trial in which Compound 1 was evaluated in patients with PBC. Compound 1 shows an overall selective activation of S1P1, S1P4, and S1P5 and the potential for safe administration to individuals with liver impairment, and thus represents a much needed option for the treatment of individuals with PBC.

SUMMARY OF THE INVENTION

In its various embodiments, the present invention is directed to, inter alia, methods of treating primary biliary cholangitis (PBC) in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof. In some embodiments, primary biliary cholangitis (PBC) is primary biliary cirrhosis.

In other embodiments, the present invention is directed to uses of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically acceptable salt, hydrate, or solvate thereof, in the manufacture of a medicament for the treatment of primary biliary cholangitis (PBC).

In some embodiments, the individual was previously treated with a therapeutically effective amount of ursodeoxycholic acid (UDCA).

In some embodiments, the individual is currently treated with a therapeutically effective amount of ursodeoxycholic acid (UDCA).

In some embodiments, the therapeutically effective amount of UDCA is substantially the same amount (stable dose) for at least 6 months.

In some embodiments, the therapeutically effective amount of UDCA is substantially the same amount (stable dose) for at least 3 months.

In some embodiments, the individual was previously treated with ursodeoxycholic acid (UDCA) and the individual had an inadequate response to UDCA.

In some embodiments, the individual had an inadequate response to UDCA as determined by an alkaline phosphate (ALP) > 1.67 x upper limit of normal (ULN) for the individual.

In some embodiments, the individual had an inadequate response after 6 months of treatment with UDCA.

In some embodiments, the individual had an inadequate response after 6 months of treatment with UDCA and an alkaline phosphate (ALP) > 1.67 x upper limit of normal (ULN).

In some embodiments, the treatment dose of UDCA was at least 13 mg/kg/day.

In some embodiments, the individual has at least one primary biliary cholangitis diagnosis criteria selected from the group consisting of:

-   anti-mitochondrial antibody (AMA) titer >1:40; -   alkaline phosphate (ALP) > 1.5 x ULN for at least 6 months; and -   liver biopsy findings consistent with PBC.

In some embodiments, the individual has at least two primary biliary cholangitis diagnosis criteria selected from the group consisting of:

-   anti-mitochondrial antibody (AMA) titer >1:40; -   alkaline phosphate (ALP) > 1.5 x ULN for at least 6 months; and -   liver biopsy findings consistent with PBC.

In some embodiments, the individual has at least one of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio(INR) < 1.2 x ULN; and -   serum creatinine < 1.5 mg/dL (133 µmol/L).

In some embodiments, the individual has at least one of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio(INR) < 1.2 x ULN; and -   serum creatinine < 1.5 mg/dL (133 µmol/L).

In some embodiments, the individual has at least one additional condition selected from the group consisting of: pruritus, fatigue, osteoporosis, vitamin deficiencies, dry eyes and/or mouth, portal hypertension, and pain.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is administered orally.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is formulated as a capsule or tablet suitable for oral administration.

In some embodiments, the individual is administered a titration dose of Compound 1 prior to the administering the therapeutically effective amount of Compound 1, wherein the titration dose is less than the therapeutically effective amount of Compound 1.

In some embodiments, the titration dose is less than about 2 mg of Compound 1.

In some embodiments, the titration dose is maintained until there are no significant changes in vital signs and/or EKG of the individual.

In some embodiments, the titration dose is maintained until the pulse rate ≥ 55 bpm, systolic blood pressure (SBP) ≥ 90, and diastolic blood pressure (DBP) ≥ 55 mmHg in the individual.

In some embodiments, the titration dose is maintained for no more than 14 days prior to administering the therapeutically effective amount of Compound 1.

In some embodiments, the titration dose comprises a first titration dose and a second titration dose, wherein the first titration dose is less than the second titration dose and each titration dose is less than the therapeutically effective amount of Compound 1.

In some embodiments, the first titration dose is in an amount equivalent to about 1 mg of Compound 1.

In some embodiments, the second titration dose is in an amount equivalent to about 1.5 mg of Compound 1.

In some embodiments, the first titration dose is in an amount equivalent to about 1 mg of Compound 1 and the second titration dose is in an amount equivalent to about 1.5 mg of Compound 1.

In some embodiments, the first titration dose is maintained for no more than 7 days prior to administering the second titration dose.

In some embodiments, the second titration dose is maintained for no more than 7 days prior to administering the therapeutically effective amount of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1.0 mg to about 5 mg of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 2 mg of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is selected from: Compound 1, a calcium salt of Compound 1, and an L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an anhydrous, non-solvated crystalline form of the L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is a crystalline free-plate habit of the non-solvated L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an anhydrous, non-solvated crystalline form of Compound 1.

In some embodiments, the method further comprises administering Compound 1, or a pharmaceutically salt, solvate, or hydrate thereof, in combination with a therapeutically effective amount of with a compound selected from the group consisting of: an antihistamine (diphenhydramine), cholestyramine (questran, prevalite), rifampin, an opioid antagonist (naloxone), pilocarpine (isopto carpine, salagen), cevimeline (evoxac), calcium and/or vitamin D supplement, and vitamin A, D, E and/or K supplement.

Some embodiments of the present invention related to titration packages for enabling compliance with a regimen of changing dosage of a medication over a period of time for the treatment of primary biliary cholangitis (PBC), wherein the medication is (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) or a pharmaceutically salt, solvate, or hydrate thereof, the package comprising:

-   a first number of daily units of a pharmaceutical composition     comprising one or more doses of Compound 1, or a pharmaceutically     acceptable salt, hydrate, or solvate thereof, wherein each dose is     in an amount equivalent to about 1.5 mg or less of Compound 1, and -   a second number of daily units of a pharmaceutical composition     comprising a standard dose of Compound 1, or a pharmaceutically     acceptable salt, hydrate, or solvate thereof, is in an amount     equivalent to about 1.0 to about 2.5 mg of Compound 1.

In some embodiments, the dose of the first number of daily units is in an amount equivalent to about 1 mg of Compound 1.

In some embodiments, the dose of the first number of daily units is in an amount equivalent to about 1.5 mg of Compound 1.

In some embodiments, the dose of the second number of daily units is in an amount equivalent to about 2 mg of Compound 1.

In some embodiments, the dose of the first number of daily units is in an amount equivalent to about 1 mg or 1.5 mg of Compound land the dose of the second number of daily units is in an amount equivalent to about 2 mg of Compound 1.

Some embodiments relate to kits comprising a titration package according to any previous embodiment, and instructions indicating that the medication is to be administered to an individual in need of treatment of primary biliary cholangitis (PBC).

These and other aspects of the invention disclosed herein will be set forth in greater detail as the patent disclosure proceeds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a PXRD Pattern overlay for the L-arginine salt of Compound 1 showing the peak intensity differences between plates and spherulites indicating a higher degree of crystallinity for the plates compared to the spherulites. Also shown is the lower sample-related background scatter (i.e., a lower amorphous halo contribution) for the plates. However, the plates and spherulites are observed to show the same crystal phase.

FIG. 2 shows a schedule of procedures and visits for screening, treatment, and follow-up periods for individuals related to the clinical study described in Example 2.

FIG. 3 shows a schedule of procedures and visits for the screening period for individuals related to the clinical study described in Example 2.

FIG. 4 shows a schedule of procedures and visits for the treatment period (Part 1) for individuals related to the clinical study described in Example 2.

FIG. 5 shows a schedule of procedures and visits for the treatment period (Part 2) for individuals related to the clinical study described in Example 2.

FIG. 6 shows a schedule of procedures and visits for the follow-up period for individuals related to the clinical study described in Example 2.

FIG. 7 shows a flowchart for the preparation of core tablets of the L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1).

Footnotes and abbreviations found in FIGS. 2 to 6 are provided below:

-   AE: adverse event; ALP: alkaline phosphatase; C4: 7     alpha-hydroxy-4-cholesten-3-one; ECG: electrocardiogram; EOS: end of     study; EOT: end of treatment; EWD: early withdrawal; FU: follow up;     HBsAg: hepatitis B surface antigen; HCV: hepatitis C virus; HIV:     human immunodeficiency virus; PK: pharmacokinetics; PML: progressive     multifocal leukoencephalopathy; pSS: primary Sjögren’s syndrome; TB:     tuberculosis. -   ^(a) For patients with dose escalation who tolerate the 1 mg dose     level based on available PK from Day 1 and Week 2, and safety data;     Week 14 visit will only take place for these dose-escalated     patients. -   ¹ Interim/abbreviated physical exam only. -   ² Blood samples for PK will be collected at up to 45 min pre-dose,     and at 2, 4, 6, and 8 hours post-dose for Day 1 and Week 2, and     optionally at 12 and/or 24 hours post-dose at Week 2. Week 12: If a     patient does not dose escalate, a pre-dose sample will be taken. If     a patient dose escalates to 2 mg daily dose, a pre-dose, 6- and     8-hour post-dose sample will be collected. Week14: If a patient is     on a 2 mg daily dose a pre-dose and 2, 4, 6, 8, and optional 12     and/or 24 hour post-dose sample will be collected. Week 24: all     patients will have a PK sample collected approximately 24 hours     after their last dose during the Week 24 visit with 2 optional     timepoints between 72 hours and 1 week after their last dose. Day 1     and Week 2 data will provide guidance for potential dose escalation     of etrasimod at week 12. -   ³ Vital signs and 12-lead ECG will be captured hourly on     Day1/baseline and Week 2 pre-dose through at least 8 hours post-dose     at the clinic. On Week 12 and Week 14, for those patients with dose     escalation, vital signs and ECG monitoring will be performed hourly     until at least 8 hours post dosing at the clinic. On Week 12, for     those patients without dose escalation, ECG monitoring will be     performed predose. For the rest of clinical visits (Weeks 4, 8, 16,     20, and 24), vital signs and 12-lead ECG will be captured at     pre-dose. 24-hour Holter monitor will be performed 24 hours before     dosing and through 24 hours post dosing on Day 1 and at Week 12 (if     dose escalation occurred). Typically, all safety ECGs will be     obtained as single tracings, with the exception of the pretreatment     ECG obtained on Day 1, which is a triplicate recording. -   ⁴ For all female patients, a urine dipstick pregnancy test is     required at Day 1 and a serum hCG test is required for all other     indicated visits. -   ⁵ For the first 12 weeks of dose titration, individual patients will     receive 1 mg q.d. if tolerable, followed by 2 mg q.d. at Week 12.     During the rest of the study, dosage should be maintained at 2 mg     q.d. -   ⁶ FSH - only in women to confirm the postmenopausal status. -   ⁷ For patients with abnormal results at screening. -   ⁸ If the absolute peripheral lymphocyte count has not recovered to     at least 80% of the baseline value, or reached normal ranges, at the     2-week follow-up, the patient must return for weekly CBC tests until     the absolute peripheral lymphocyte count has returned to at least     these values. AE: adverse event; ALP: alkaline phosphatase; C4: 7     alpha-hydroxy-4-cholesten-3-one; ECG: electrocardiogram; EOS: end of     study; EOT: end of treatment; EWD: early withdrawal; FU: Follow up;     HBsAg: hepatitis B surface antigen; HCV: hepatitis C virus; HIV:     human immunodeficiency virus; PK: pharmacokinetics; PML: progressive     multifocal leukoencephalopathy; pSS: primary Sjogren’s syndrome; TB:     tuberculosis.

DETAILED DESCRIPTION OF THE INVENTION

The present disclosure provides, inter alia, methods of treating primary biliary cholangitis (PBC) in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

In other embodiments, the present invention is directed to uses of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically acceptable salt, hydrate, or solvate thereof, in the manufacture of a medicament for the treatment of primary biliary cholangitis (PBC) in an individual.

In other embodiments, the present invention is directed to (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically acceptable salt, hydrate, or solvate thereof, for use in the treatment of primary biliary cholangitis (PBC).

In other embodiments, the present invention is directed to titration packages for enabling compliance with a regimen of changing dosage of a medication over a period of time for the treatment of primary biliary cholangitis (PBC), wherein the medication is (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) or a pharmaceutically salt, solvate, or hydrate thereof, the package comprising:

-   a first number of daily units of a pharmaceutical composition     comprising one or more doses of Compound 1, or a pharmaceutically     acceptable salt, hydrate, or solvate thereof, wherein each dose is     in an amount equivalent to about 1.5 mg or less of Compound 1, and -   a second number of daily units of a pharmaceutical composition     comprising a standard dose of Compound 1, or a pharmaceutically     acceptable salt, hydrate, or solvate thereof, is in an amount     equivalent to about 1.0 to about 2.5 mg of Compound 1.

Certain processes for the preparation of Compound 1 and the L-arginine salt of Compound 1 have been previously described; see WO2010/011316 and WO2011/094008. In addition, the novel crystalline plate habit for the L-arginine salt of Compound 1 has been previously described and is referred to herein as, “crystalline free-plate habit of the non-solvated L-arginine salt of Compound 1”; see WO2016/209809.

In some embodiments, the methods provided herein are for the treatment of primary biliary cholangitis that has progressed to primary biliary cirrhosis. In some embodiments, the methods provided herein are for the prevention of primary biliary cirrhosis. In some embodiments, the methods provided herein are for delayed progression to primary biliary cirrhosis.

The following is a list of abbreviations: ACS (acute coronary syndrome); ADL (activities of daily living); AE (adverse event); ALP (alkaline phosphatase); ALT (alanine aminotransferase (SGPT)); AMA (anti-mitochondrial antibodies); ANA (anti-nuclear antibodies); AST (aspartate aminotransferase (SGOT)); AV (atrio-ventricular); bpm (beats per minute); CBC (complete blood count); CFR (Code of Federal Regulations); CI (confidence interval); CRF (case report form); CRP (C-reactive protein); CRO (contract research organization); D (day); DILI (drug-induced liver injury); ECG (electrocardiogram); ED50 (half maximal dose); eGFR (estimated glomerular filtration rate); ELISA (enzyme-linked immunosorbent assay); EOS (end of study); EOT (end of treatment); FDA (Food and Drug Administration); FEV1 (forced expiratory volume ); FU (follow up); FVC (forced vital capacity); GCP (Good Clinical Practice); GGT (gamma glutamyl transferase); HBsAg (hepatitis B surface antigen); hCG (human chorionic gonadotropin); HCV (hepatitis C virus); HREC (human research ethics committee (AUS)); HIV (human immunodeficiency virus); HR (heart rate); ICH (International Conference on Harmonization); ICF (informed consent form); IEC (Independent Ethics Committee); IND (Investigational New Drug); IRB (Institutional Review Board); INR (international normalized ratio); hormone-releasing system); IUD (intrauterine device); IUS (Intrauterine hormone-releasing system); kg (kilogram); LDH (lactate dehydrogenase); L (liter); MCH (mean corpuscular hemoglobin); MCV (mean corpuscular volume); MedDRA (Medical Dictionary for Regulatory Activities); mg (milligram); MI (myocardial infarction); NK (natural killer); NOAEL (no observed adverse effect level); OTC (over-the -counter); PBC (peripheral biliary cholangitis); PBL (peripheral blood lymphocyte); PD (pharmacodynamics); PFT (pulmonary function test); PGA (Physicians Global Assessments); PI (Principal Investigator); PK (pharmacokinetics); p.o. (per os (orally)); PRO (patient reported outcome); pSS (primary Sjögren’s syndrome); PVG (pharmacovigilance); q.d. (quaque die (once daily)); SAP (statistical analysis plan); S1P (sphingosine 1-phosphate); SAE (serious adverse event); SBP (systolic blood pressure); SD (standard deviation); sec (second); SOP(s) (standard operating procedure(s)); t1/2 (elimination half-life); tmax (the median time to reach maximum plasma concentration); TBUT (Tear Film Break-up Time ); TIA (transient ischemic attack); UDCA (ursodeoxycholic acid); ULN (upper limit of normal); VS (vital signs); VZV (varicella zoster virus); WBC (white blood cell); WHO (World Health Organization); and WHODRUG (World Health Organization Drug Dictionary).

Crystalline L-Arginine Salt of Compound 1

The crystalline free-plate habit or morphology and processes useful in the preparation of a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)-benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid are described in WO2016/209809. The plates were discovered from the novel synthetic methods and were shown to be thin hexagonal-like plates with two opposite sides of the plate being longer that the other sides (i.e., elongated hexagonal plate). However, due to the thin characteristic of the plates, a complete unbroken plate is rarely seen. Instead, what is generally observed are large to small broken pieces of the thin hexagonal-like plates. It is understood by those skilled in the art that microscopy is one of the more useful techniques to distinguish two crystalline habits or morphologies. This is particularly useful when 2 or more morphologies are associated with the same or substantially the same crystal phase as is the case with the L-arginine salt of Compound 1. Comparing the PXRD patterns of the habit prepared previously (i.e., WO2011/094008) and the plate habit prepared as described in WO2016/209809 (i.e., see FIG. 1 , PXRD overlay between spherulites and plates) it was observed that the two PXRD patterns were the same or substantially the same, thus the two habits represented the same crystal phase.

Although the two habits revealed the same or substantially the same PXRD pattern, a higher degree of crystallinity was observed for the plate habit as indicated by substantially higher peak intensities and yet lower sample-related background scatter (i.e., a lower amorphous halo contribution). Since sample size and sample preparation can affect peak intensities and sample-related background scatter, and since the two habits share the same crystal phase, PXRD may not be considered the most appropriate test method to distinguish between two habits. However, PXRD does allow for determining whether two habits have the same crystal phase or different crystal phases. For determining different habits, microscopy is one of the more useful methods. Accordingly, the skilled person would be capable of reviewing a micrograph for a crystal habit and readily determine the crystal habit.

In addition to the techniques recognized in the art, specific surface can also be used to characterize a habit, such as the free-plates. Accordingly, the specific surface area values disclosed in the present invention have been obtained by means of a specific surface area analysis technique based on the BET (Brunauer, Emmett and Teller) theory, which is a well-accepted theory known in the art for the calculation of surface areas of solids by means of measuring their physical adsorption of gas molecules (see: Brunauer, S.; Emmett, P. H.; and Teller, E.; J. Am. Chem. Soc., 1938, 60, 309). In particular, the specific surface area values measured in the present invention have been calculated from the BET surface area plot obtained by measuring the quantity of nitrogen gas molecules adsorbed by a weighted amount of solid at different relative pressures (P/Po) within the range 0.05-0.3 (P/P₀), at 77.3 K. The measurement of the adsorption of gas molecules was carried out by means of a Micromeritics™ TriStar II BET surface analyzer having the characteristics as set out below in Example 3. Namely, nitrogen gas was used for the adsorption measurement. The sample for each analysis was degassed at 25° C. for 960 minutes under vacuum (i.e., 100 mm/Hg). The determination of the adsorption of nitrogen was measured at 77.3 K at eleven relative pressures (P/P₀) sufficiently dispersed within the range of about 0.05 to about 0.30 (i.e. eleven absolute pressures in the range of about 36 mm Hg to about 223 mm Hg relative to the saturated pressure at the time of measurement that ranged from about 738 mmHg to about 743 mmHg).

One aspect of the present invention relates to a novel crystalline plate morphology of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid as described herein.

One aspect of the present invention relates to a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid.

In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, and 20.5° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, and 28.8° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8° ± 0.2°, and 37.3° ± 0.2°.

In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.0° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.0° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.5° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 206.0° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 206.5° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.5° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 206.5° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 207.0° C. to 208.1° C. at a scan rate of 10° C./minute.

In some embodiments, the crystalline free-plate habit has a dynamic moisture sorption (DMS) profile with an adsorption phase from 30% RH to 90% RH wherein the crystalline free-plate habit gains about 0.3% weight or less at 90% RH. In some embodiments, the crystalline free-plate habit has a dynamic moisture sorption (DMS) profile with an adsorption phase from 30% RH to 90% RH wherein the crystalline free-plate habit gains about 0.2% weight or less at 90% RH.

One aspect of the present invention relates to a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid having a BET specific surface area of about 0.05 m²/g, about 0.1 m²/g, about 0.15 m²/g, about 0.2 m²/g, about 0.25 m²/g, about 0.3 m²/g, about 0.35 m²/g, about 0.4 m²/g, about 0.45 m²/g, about 0.5 m²/g, about 0.55 m²/g, about 0.6 m²/g, about 0.65 m²/g, or about 0.7 m²/g to about 2.0 m²/g, about 2.5 m²/g, about 3.0 m²/g, about 3.5 m²/g, about 4.0 m²/g, about 4.5 m²/g, about 5.0 m²/g, about 5.5 m²/g, about 6.0 m²/g, about 6.5 m²/g, about 7.0 m²/g, about 7.5 m²/g, about 8.0 m²/g, about 8.5 m²/g, about 9.0 m²/g, or about 9.5 m²/g.

In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.1 m²/g to about 5.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.1 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.3 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.5 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.6 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.3 m²/g to about 3.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.4 m²/g to about 2.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.5 m²/g to about 1.8 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.6 m²/g to about 1.6 m²/g.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8°     ± 0.2°, and 37.3° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.0° C. to 208.5° C. at     a scan rate of 10° C./minute; and/or -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.3% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 206.5° C. to t 208.5° C.     at a scan rate of 10° C./minute; and/or -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.3% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.5° C. to 208.5° C. at     a scan rate of 10° C./minute; and/or -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.2% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° + 0.2°, and 20.5° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 207.1° C. to 208.1° C. at     a scan rate of 10° C./minute; and/or -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.2% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.0° C. to 208.5° C. at     a scan rate of 10° C./minute; and/or -   2) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.3% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 206.5° C. to t 208.5° C.     at a scan rate of 10° C./minute; and/or -   2) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.3% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.5° C. to 208.5° C. at     a scan rate of 10° C./minute; and/or -   2) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.2% weight or less at 90% RH.

In some embodiments, the crystalline free-plate habit has:

-   1) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 207.1° C. to 208.1° C. at     a scan rate of 10° C./minute; and/or -   2) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein the crystalline free-plate habit     gains about 0.2% weight or less at 90% RH. In some embodiments, the     crystalline free-plate habit has: -   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, and 20.5° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.0° C. to 208.5° C. at     a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.3% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.1 m²/g to about 5.0 m²/g.     In some embodiments, the crystalline free-plate habit has: -   1) a powder X-ray dittraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.5° C. to t 208.5° C.     at a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.3% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.1 m²/g to about 4.0 m²/g.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.5° C. to t 208.5° C.     at a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.3% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.3 m²/g to about 3.0 m²/g.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.5° C. to 208.5° C. at     a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.3% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.6 m²/g to about 4.0 m²/g.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 206.5° C. to t 208.5° C.     at a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.3% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.4 m²/g to about 2.0 m²/g.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, and     28.8° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 206.5° C. to 208.1° C. at     a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.2% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.5 m²/g to about 1.8 m²/g.

In some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8°     ± 0.2°, and 37.3° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 205.5° C. to 208.1° C. at     a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.2% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.6 m2/g to about 4.0 m2/g.

in some embodiments, the crystalline free-plate habit has:

-   1) a powder X-ray diffraction pattern comprising peaks, in terms of     2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8°     ± 0.2°, and 37.3° ± 0.2°; -   2) a differential scanning calorimetry trace comprising an endotherm     with an extrapolated onset temperature of 207.1° C. to 208.1° C. at     a scan rate of 10° C./minute; -   3) a dynamic moisture sorption (DMS) profile with an adsorption     phase from 30% RH to 90% RH wherein said crystalline free-plate     habit gains about 0.2% weight or less at 90% RH; and/or -   4) a BET specific surface area of about 0.6 m²/g to about 1.6 m²/g.

One aspect of the present invention relates to a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid having a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, and 20.5° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, and 28.8° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8° ± 0.2°, and 37.3° ± 0.2°.

One aspect of the present invention relates to a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid having a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.0° C. to 208.5° C. when scanned at 10° C. per minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.0° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.5° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 205.5° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 206.0° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 206.5° C. to 208.5° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 206.5° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a differential scanning calorimetry trace comprising an endotherm with an extrapolated onset temperature of 207.0° C. to 208.1° C. at a scan rate of 10° C./minute. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, and 20.5° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, and 28.8° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8° ± 0.2°, and 37.3° ± 0.2°.

One aspect of the present invention relates to a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid having a dynamic moisture sorption (DMS) profile with an adsorption phase from 30% RH to 90% RH wherein the crystalline free-plate habit gains about 0.3% weight or less at 90% RH. In some embodiments, the crystalline free-plate habit has a dynamic moisture sorption (DMS) profile with an adsorption phase from 30% RH to 90% RH wherein the crystalline free-plate habit gains about 0.2% weight or less at 90% RH. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, and 20.5° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, and 28.8° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8° ± 0.2°, and 37.3° ± 0.2°.

One aspect of the present invention relates to a crystalline free-plate habit of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid having a BET specific surface area of about 0.1 m²/g to about 5.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.1 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.3 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.5 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.6 m²/g to about 4.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.3 m²/g to about 3.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.4 m²/g to about 2.0 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.5 m²/g to about 1.8 m²/g. In some embodiments, the crystalline free-plate habit has a BET specific surface area of about 0.6 m²/g to about 1.6 m²/g. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, and 20.5° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, and 24.6° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, and 28.8° ± 0.2°. In some embodiments, the crystalline free-plate habit has a powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at 8.2° ± 0.2°, 16.4° ± 0.2°, 20.5° ± 0.2°, 24.6° ± 0.2°, 28.8° ± 0.2°, and 37.3° ± 0.2°.

Certain Embodiments

In its various embodiments, the present invention is directed to, inter alia, methods of treating primary biliary cholangitis (PBC) in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopcntyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

In other embodiments, the present invention is directed to uses of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically acceptable salt, hydrate, or solvate thereof, in the manufacture of a medicament for the treatment of primary biliary cholangitis (PBC) in an individual.

In some embodiments, the individual was previously treated with a therapeutically effective amount of ursodeoxycholic acid (UDCA).

In some embodiments, the individual is currently treated with a therapeutically effective amount of ursodeoxycholic acid (UDCA).

In some embodiments, the therapeutically effective amount of UDCA is about 10-20 mg/kg/day. In some embodiments, the therapeutically effective amount of UDCA is about 13-15 mg/kg/day.

In some embodiments, the therapeutically effective amount of UDCA is reduced when administered with Compound 1. In some embodiments, the therapeutically effective amount of UDCA is reduced to less than 15 mg/kg/day. In some embodiments, the therapeutically effective amount of UDCA is reduced to less than 13 mg/kg/day. In some embodiments, the therapeutically effective amount of UDCA is reduced to less than 500 mg. In some embodiments, the therapeutically effective amount of UDCA is reduced to less than 500 mg per administration. In some embodiments, the therapeutically effective amount of UDCA is reduced to less than 250 mg. In some embodiments, the therapeutically effective amount of UDCA is reduced to less than 250 mg per administration In some embodiments, the frequency of administration of UDCA is reduced. In some embodiments, the administration of UDCA is reduced to four divided doses. In some embodiments, the administration of UDCA is reduced to three divided doses. In some embodiments, the administration of UDCA is reduced to two divided doses. In some embodiments, UDCA is administered four times daily. In some embodiments, UDCA is administered three times daily. In some embodiments, UDCA is administered twice daily.

In some embodiments, the therapeutically effective amount of Compound 1 is reduced when administered with UDCA. In some embodiments, the therapeutically effective amount of Compound 1 is reduced to less than 2 mg. In some embodiments, the therapeutically effective amount of Compound 1 is reduced to less than 1.5 mg. In some embodiments, the therapeutically effective amount of Compound 1 is reduced to less than 1 mg. In some embodiments, the frequency of administration of Compound 1 is reduced. In some embodiments, Compound 1 is administered once daily. In some embodiments, Compound 1 is administered every other day.

In some embodiments, the therapeutically effective amount of UDCA is about 100 mg to about 1000 mg. In some embodiments, the therapeutically effective amount of UDCA is about 250 mg to about 500 mg. In some embodiments, the therapeutically effective amount of UDCA is about 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 925 mg, 950 mg, 975 mg, or 1000 mg. In some embodiments, the therapeutically effective amount of UDCA is about 250 mg. In some embodiments, the therapeutically effective amount of UDCA is about 500 mg.

In some embodiments, the therapeutically effective amount of UDCA is administered in two to ten divided doses. In some embodiments, the therapeutically effective amount of UDCA is administered in two to four divided doses. In some embodiments, the therapeutically effective amount of UDCA is administered in two divided doses. In some embodiments, the therapeutically effective amount of UDCA is administered in three divided doses. In some embodiments, the therapeutically effective amount of UDCA is administered in four divided doses. In some embodiments, the therapeutically effective amount of UDCA is administered in five divided doses. In some embodiments, UDCA is administered four times daily. In some embodiments, UDCA is administered three times daily. In some embodiments, UDCA is administered twice daily.

In some embodiments, the therapeutically effective amount of UDCA is substantially the same amount (stable dose) for at least 6 months.

In some embodiments, the individual was previously treated with ursodeoxycholic acid (UDCA) and the individual had an inadequate response to UDCA.

In some embodiments, the individual had an inadequate response to UDCA as determined by an alkaline phosphate (ALP) > 1.67 x upper limit of normal (ULN) for the individual.

In some embodiments, the individual had an inadequate response after 6 months of treatment with UDCA.

In some embodiments, the individual had an inadequate response after 6 months of treatment with UDCA and an alkaline phosphate (ALP) > 1.67 x upper limit of normal (ULN).

In some embodiments, the treatment dose of UDCA was at least 13 mg/kg/day.

In some embodiments, the individual has at least one primary biliary cholangitis diagnosis criteria selected from the group consisting of:

-   anti-mitochondrial antibody (AMA) titer >1:40; -   alkaline phosphate (ALP) >1.5 x ULN for at least 6 months; and -   liver biopsy findings consistent with PBC.

In some embodiments, the individual has at least two primary biliary cholangitis diagnosis criteria selected from the group consisting of:

-   anti-mitochondrial antibody (AMA) titer >1:40; -   alkaline phosphate (ALP) > 1.5 x ULN for at least 6 months; and -   liver biopsy findings consistent with PBC.

In some embodiments, the individual has at least one of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but <10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; and -   serum creatinine < 1.5 mg/dL (133 µmol/L).

In some embodiments, the individual has at least one of the criteria selected from the group consisting of:

-   ALP >1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; and -   serum creatinine < 1.5 mg/dL (133 µmol/L).

In some embodiments, the individual has at least one of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least one of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan® (transient elastography) < 10 kPa.

In some embodiments, the individual has at least two of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least two of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least three of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least three of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least four of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least four of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least five of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least five of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum ciealinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least six of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least six of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least seven of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least seven of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least eight of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least eight of the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has all the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < 1.5 x ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.4 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has all the criteria selected from the group consisting of:

-   ALP > 1.67 x ULN but < 10 x ULN; -   ALT and AST < 5 x ULN; -   total bilirubin < ULN; -   international normalized ratio (INR) < 1.2 x ULN; -   platelet count > 150,000 /mm³; -   serum albumin > 3.0 g/dL; -   serum creatinine < 1.5 mg/dL (133 µmol/L); -   TSH < 5.0 mU/L without clinical significant changes of free T3 and     T4 levels; and -   Fibroscan (transient elastography) < 10 kPa.

In some embodiments, the individual has at least one additional condition selected from the group consisting of: pruritus, fatigue, osteoporosis, vitamin deficiencies, dry eyes and/or mouth, portal hypertension, and pain.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is administered orally.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is formulated as a capsule or tablet suitable for oral administration.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is formulated as a tablet suitable for oral administration.

In some embodiments, the individual is administered a titration dose of Compound 1 prior to the administering the therapeutically effective amount of Compound 1, wherein the titration dose is less than the therapeutically effective amount of Compound 1.

In some embodiments, the titration dose is less than about 2 mg of Compound 1.

In some embodiments, the titration dose is maintained until there are no significant changes in vital signs and/or EKG of the individual.

In some embodiments, the titration dose is maintained until the pulse rate ≥ 55 bpm, systolic blood pressure (SBP) ≥ 90, and diastolic blood pressure (DBP) ≥ 55 mmHg in the individual.

In some embodiments, the titration dose is maintained for no more than 14 days prior to administering the therapeutically effective amount of Compound 1.

In some embodiments, the titration dose comprises a first titration dose and a second titration dose, wherein the first titration dose is less than the second titration dose and each titration dose is less than the therapeutically effective amount of Compound 1.

In some embodiments, the first titration dose is in an amount equivalent to about 1 mg of Compound 1.

In some embodiments, the second titration dose is in an amount equivalent to about 1.5 mg of Compound 1.

In some embodiments, the first titration dose is in an amount equivalent to about 1 mg of Compound 1 and the second titration dose is in an amount equivalent to about 1.5 mg of Compound 1.

In some embodiments, the first titration dose is maintained for no more than 7 days prior to administering the second titration dose.

In some embodiments, the second titration dose is maintained for no more than 7 days prior to administering the therapeutically effective amount of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1.0 mg to about 5 mg of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg to about 2 mg.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75, or about 2 mg.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg or about 2 mg.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, or 2 mg of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to ≥ 1 mg of Compound 1. In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to ≥ 1.5 mg of Compound 1. In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to ≤ 1 mg of Compound 1. In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to ≤ 1.5 mg of Compound 1. In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to ≤ 2 mg of Compound 1. In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is no more than 1 mg, 1.5 mg, 2 mg, or 5 mg of Compound 1.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is administered once daily.

In some embodiments, the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 2 mg of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is selected from: Compound 1, a calcium salt of Compound 1, and an L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an anhydrous, non-solvated crystalline form of the L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is a crystalline free-plate habit of the non-solvated L-arginine salt of Compound 1.

In some embodiments, the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an anhydrous, non-solvated crystalline form of Compound 1.

In some embodiments, the method further comprises administering Compound 1, or a pharmaceutically salt, solvate, or hydrate thereof, in combination with a therapeutically effective amount of with a compound selected from the group consisting of: an antihistamine (diphenhydramine), cholestyramine (questran, prevalite), rifampin, an opioid antagonist (naloxone), pilocarpine (isopto carpine, salagen), cevimeline (evoxac), calcium and/or vitamin D supplement, and vitamin A, D, E and/or K supplement.

Some embodiments of the present invention related to titration packages for enabling compliance with a regimen of changing dosage of a medication over a period of time for the treatment of primary biliary cholangitis (PBC), wherein the medication is (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) or a pharmaceutically salt, solvate, or hydrate thereof, the package comprising:

-   a first number of daily units of a pharmaceutical composition     comprising one or more doses of Compound 1, or a pharmaceutically     acceptable salt, hydrate, or solvate thereof, wherein each dose is     in an amount equivalent to about 1.5 mg or less of Compound 1, and -   a second number of daily units of a pharmaceutical composition     comprising a standard dose of Compound 1, or a pharmaceutically     acceptable salt, hydrate, or solvate thereof, is in an amount     equivalent to about 1.0 to about 2.5 mg of Compound 1.

In some embodiments, the dose of the first number of daily units is in an amount equivalent to about 1 mg of Compound 1.

In some embodiments, the dose of the first number of daily units is in an amount equivalent to about 1.5 mg of Compound 1.

In some embodiments, the dose of the second number of daily units is in an amount equivalent to about 2 mg of Compound 1.

In some embodiments, the dose of the first number of daily units is in an amount equivalent to about 1 mg or 1.5 mg of Compound 1and the dose of the second number of daily units is in an amount equivalent to about 2 mg of Compound 1.

Some embodiments relate to kits comprising a titration package according to any previous embodiment, and instructions indicating that the medication is to be administered to an individual in need of treatment of primary biliary cholangitis (PBC).

In some embodiments, the individual is administered the therapeutically effective amount of Compound 1 once daily.

In some embodiments, the individual is administered the therapeutically effective amount of Compound 1 twice daily.

In some embodiments, the individual is administered the therapeutically effective amount of Compound 1 three times daily.

In some embodiments, the individual is administered the therapeutically effective amount of Compound 1 once every other day.

In some embodiments, Compound 1 is administered without food.

In some embodiments, the individual has been identified as having impaired liver function prior to administration of the titration dose.

Some embodiments relate to methods, further comprising:

-   identifying the liver function of the individual; and -   administering at least one titration dose if the individual has     impaired liver function, or -   not administering a titration dose if the individual does not have     impaired liver function.

Some embodiments relate to methods, wherein impaired liver function of the individual is determined by a liver function test for at least one marker selected from: bilirubin, albumin, total protein, aminotransferase (ALT), aspartate aminotransferase (AST), creatinine kinase (CK), gamma-glutamyl transferase (GGT), or alkaline phosphatase (ALP).

One aspect of the present invention relates to methods of treating an individual in need thereof with (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) comprising:

-   (a) analyzing one or more samples from the individual for the level     of at least one biomarker obtained prior to the treatment with     Compound 1; and -   (b) administering Compound 1 or not administering Compound 1 to the     individual based on a predetermined level of the at least one     biomarker prior to treatment of Compound 1; -   wherein the at least one biomarker is selected from the group     consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high     sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase     (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl     transferase (GGT); (vii) anti mitochondrial antibodies (AMA); (viii)     Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4     (C4); (xi) IgG; and (xii) IgM.

One aspect of the present invention relates to methods of treating an individual in need thereof with (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) comprising:

-   (a) administering Compound 1 to the individual; -   (b) analyzing one or more samples from the individual for the level     of at least one biomarker obtained after the treatment with Compound     1; and -   (c) (i) continuing administration of Compound 1 if the at least one     biomarker is less than or equal to a predetermined level for the at     least one biomarker prior to treatment of Compound 1; or     -   (ii) discontinuing administration of Compound 1 if the at least         one biomarker is greater than a predetermined level for the at         least one biomarker prior to treatment of Compound 1; -   wherein the at least one biomarker is selected from the group     consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high     sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase     (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl     transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii)     Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4     (C4); (xi) IgG; and (xii) IgM.

One aspect of the present invention relates to methods of treating an individual in need thereof with (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) comprising:

-   (a) analyzing one or more samples from the individual for a first     level of at least one biomarker obtained prior to the treatment with     Compound 1; -   (b) administering Compound 1 to the individual; -   (c) analyzing one or more samples from the individual for a second     level of the at least one biomarker obtained after the treatment     with Compound 1; and -   (d) (i) continuing administration of Compound 1 if the second level     of the at least one biomarker in step (c) is less than or about     equal to the corresponding first level of the at least one biomarker     in step (a); or     -   (ii) discontinuing administration of Compound 1 if the second         level of the at least one biomarker in step (c) is greater than         the corresponding first level of the at least one biomarker in         step (a); -   wherein the at least one biomarker is selected from the group     consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high     sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase     (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl     transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii)     Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4     (C4); (xi) IgG; and (xii) IgM.

In some embodiments, the individual has primary biliary cholangitis (PBC).

In some embodiments, the individual has fatigue, pruritus, eye dryness, and/or Sjogren’s syndrome (SS).

In some embodiments, the individual has fatigue.

In some embodiments, the individual has pruritus.

In some embodiments, the individual has eye dryness.

In some embodiments, the individual has Sjögren’s syndrome (SS).

In some embodiments, the second level of the at least one biomarker in step (c) is 2% less, 5% less, 10% less, 12% less, 15% less, 17% less, 20% less, 22% less, 25% less, 30% less, 35% less, 40% less, 45% less, 50% less, or > 50% less than the corresponding first level of the at least one biomarker in step (a).

In some embodiments, the at least one biomarker is two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve biomarkers selected from the group consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM.

In some embodiments, the at least one biomarker in step (c) is two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve biomarkers selected from the group consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM.

One aspect of the present invention relates to methods of treating fatigue in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating fatigue in an individual with primary biliary cholangitis (PBC) comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating pruritus in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating pruritus in an individual with primary biliary cholangitis (PBC) comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating eye dryness in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating eye dryness in an individual with primary biliary cholangitis (PBC) comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating Sjogren’s syndrome (SS) in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating Sjogren’s syndrome (SS) in an individual with primary biliary cholangitis (PBC) comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof.

One aspect of the present invention relates to methods of treating an individual in need thereof with (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) comprising:

-   (a) analyzing one or more samples from the individual for a level of     at least one biomarker obtained prior to the treatment with Compound     1; and -   (d) modifying administration of Compound 1, -   wherein the at least one biomarker is selected from the group     consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high     sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase     (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl     transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii)     Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4     (C4); (xi) IgG; and (xii) IgM.

In some embodiments, modifying the administration of Compound 1 comprises increasing the amount of Compound 1.

In some embodiments, modifying the administration of Compound 1 comprises decreasing the amount of Compound 1.

In some embodiments, the individual has been administered Compound 1 prior to modified administration.

In some embodiments, the amount of UDCA is about 13 mg/kg/day to about 15 mg/kg/day.

In some embodiments, the UDCA is administered in two to four divided doses.

In some embodiments, the amount of UDCA is about 250 mg or about 500 mg.

In some embodiments, the amount or frequency of administration of UDCA is reduced when administered with Compound 1.

In some embodiments, the amount or frequency of administration of Compound 1 is reduced when administered with UDCA.

Pharmaceutical Compositions

A further aspect of the present invention pertains to pharmaceutical compositions comprising one or more compounds as described herein and one or more pharmaceutically acceptable carriers. Some embodiments pertain to pharmaceutical compositions comprising a compound of the present invention and a pharmaceutically acceptable carrier.

Some embodiments of the present invention include a method of producing a pharmaceutical composition comprising admixing at least one compound according to any of the compound embodiments disclosed herein and a pharmaceutically acceptable carrier.

Formulations may be prepared by any suitable method, typically by uniformly mixing the active compound(s) with liquids or finely divided solid carriers, or both, in the required proportions and then, if necessary, forming the resulting mixture into a desired shape.

Conventional excipients, such as binding agents, fillers, acceptable wetting agents, tabletting lubricants and disintegrants may be used in tablets and capsules for oral administration. Liquid preparations for oral administration may be in the form of solutions, emulsions, aqueous or oily suspensions and syrups. Alternatively, the oral preparations may be in the form of dry powder that can be reconstituted with water or another suitable liquid vehicle before use. Additional additives such as suspending or emulsifying agents, non-aqueous vehicles (including edible oils), preservatives and flavorings and colorants may be added to the liquid preparations. Parenteral dosage forms may be prepared by dissolving the compound of the invention in a suitable liquid vehicle and filter sterilizing the solution before filling and sealing an appropriate vial or ampule. These are just a few examples of the many appropriate methods well known in the art for preparing dosage forms.

A compound of the present invention can be formulated into pharmaceutical compositions using techniques well known to those in the art. Suitable pharmaceutically acceptable carriers, outside those mentioned herein, are known in the art; for example, see Remington, The Science and Practice of Pharmacy, 20^(th) Edition, 2000, Lippincott Williams & Wilkins, (Editors: Gennaro et al.)

While it is possible that, for use in the prophylaxis or treatment, a compound of the invention may, in an alternative use, be administered as a raw or pure chemical, it is preferable however to present the compound or active ingredient as a pharmaceutical formulation or composition further comprising a pharmaceutically acceptable carrier.

The invention thus further provides pharmaceutical formulations comprising a compound of the invention or a pharmaceutically acceptable salt, solvate, hydrate or derivative thereof together with one or more pharmaceutically acceptable carriers thereof and/or prophylactic ingredients. The carrier(s) must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not overly deleterious to the recipient thereof.

Pharmaceutical formulations include those suitable for oral, rectal, nasal, topical (including buccal and sub-lingual), vaginal or parenteral (including intramuscular, sub-cutaneous and intravenous) administration or in a form suitable for administration by inhalation, insufflation or by a transdermal patch. Transdermal patches dispense a drug at a controlled rate by presenting the drug for absorption in an efficient manner with a minimum of degradation of the drug. Typically, transdermal patches comprise an impermeable backing layer, a single pressure sensitive adhesive and a removable protective layer with a release liner. One of ordinary skill in the art will understand and appreciate the techniques appropriate for manufacturing a desired efficacious transdermal patch based upon the needs of the artisan.

The compounds of the invention, together with a conventional adjuvant, carrier, or diluent, may thus be placed into the form of pharmaceutical formulations and unit dosages thereof and in such form may be employed as solids, such as tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs, gels or capsules filled with the same, all for oral use; in the form of suppositories for rectal administration; or in the form of sterile injectable solutions for parenteral (including subcutaneous) use. Such pharmaceutical compositions and unit dosage forms thereof may comprise conventional ingredients in conventional proportions, with or without additional active compounds or principles and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed.

For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet, capsule, suspension or liquid. The pharmaceutical composition is preferably made in the form of a dosage unit containing a particular amount of the active ingredient. Examples of such dosage units are capsules, tablets, powders, granules or suspensions, with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators such as corn starch, potato starch or sodium carboxymethylcellulose; and with lubricants such as talc or magnesium stearate. The active ingredient may also be administered by injection as a composition wherein, for example, saline, dextrose or water may be used as a suitable pharmaceutically acceptable carrier.

Compounds of the present invention or a salt, solvate, hydrate or physiologically functional derivative thereof can be used as active ingredients in pharmaceutical compositions, specifically as S1P1 receptor modulators. The term “active ingredient” is defined in the context of a “pharmaceutical composition” and is intended to mean a component of a pharmaceutical composition that provides the primary pharmacological effect, as opposed to an “inactive ingredient” which would generally be recognized as providing no pharmaceutical benefit.

The dose when using the compounds of the present invention can vary and as is customary and known to the physician, it is to be tailored to the individual conditions in each individual case. It depends, for example, on the nature and severity of the illness to be treated, on the condition of the individual, or on whether an acute or chronic disease state is treated or prophylaxis is conducted or on whether further active compounds are administered in addition to the compounds of the present invention. Representative doses of the present invention include, but are not limited to, about 1 mg to about 5 mg, about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.25 mg, about 3.5 mg, about 3.75 mg, about 4 mg, about 4.25 mg, about 4.5 mg, about 4.75 mg, and about 5 mg. Multiple doses may be administered during the day, especially when relatively large amounts are deemed to be needed, for example 2, 3 or 4 doses. Depending on the individual and as deemed appropriate by the individual’s physician or caregiver it may be necessary to deviate upward or downward from the doses described herein.

The amount of active ingredient or an active salt, solvate or hydrate derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the individual and will ultimately be at the discretion of the attendant physician or clinician. Representative factors include the type, age, weight, sex, diet and medical condition of the individual, the severity of the disease, the route of administration, pharmacological considerations such as the activity, efficacy, pharmacokinetic and toxicology profiles of the particular compound employed, whether a drug delivery system is utilized, whether an acute or chronic disease state is being treated or prophylaxis is conducted or whether further active compounds are administered in addition to the compounds of the present invention and as part of a drug combination. The dosage regimen for treating a disease condition with the compounds and/or compositions of this invention is selected in accordance with a variety factors including those cited above. Thus, the actual dosage regimen employed may vary widely and therefore may deviate from a preferred dosage regimen and one skilled in the art will recognize that dosage and dosage regimens outside these typical ranges can be tested and, where appropriate, may be used in the methods of this invention.

The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as 2, 3, 4 or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations. The daily dose can be divided, especially when relatively large amounts are administered as deemed appropriate, into several, for example 2, 3 or 4 part administrations. If appropriate, depending on individual behavior, it may be necessary to deviate upward or downward from the daily dose indicated.

For preparing pharmaceutical compositions from the compounds of the present invention, the suitable pharmaceutically acceptable carrier can be either solid, liquid or a mixture of both. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or encapsulating materials.

In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component.

In tablets, the active component is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted to the desired shape and size.

The powders and tablets may contain varying percentage amounts of the active compound. A representative amount in a powder or tablet may be from 0.5 to about 90 percent of the active compound. However, an artisan would know when amounts outside of this range are necessary. Suitable carriers for powders and tablets include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter and the like. The term “preparation” is intended to include the formulation of the active compound with encapsulating material as carrier providing a capsule in which the active component, with or without carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets and lozenges can be used as solid forms suitable for oral administration.

For preparing suppositories, a low melting wax, such as an admixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein (e.g., by stirring). The molten homogenous mixture is then poured into convenient sized molds, allowed to cool and thereby to solidify.

Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

Liquid form preparations include solutions, suspensions and emulsions, for example, water or water-propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated as solutions in aqueous polyethylene glycol solution. Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds according to the present invention may thus be formulated for parenteral administration (e.g. by injection, for example bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The pharmaceutical compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use.

Aqueous formulations suitable for oral use can be prepared by dissolving or suspending the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents, as desired.

Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents and the like.

For topical administration to the epidermis the compounds according to the invention may be formulated as ointments, creams or lotions, or as a transdermal patch.

Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents.

Formulations suitable for topical administration in the mouth include lozenges comprising the active agent in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

Solutions or suspensions are applied directly to the nasal cavity by conventional means, for example with a dropper, pipette or spray. The formulations may be provided in single or multi-dose form. In the latter case of a dropper or pipette, this may be achieved by the individual administering an appropriate, predetermined volume of the solution or suspension. In the case of a spray, this may be achieved for example by means of a metering atomizing spray pump.

Administration to the respiratory tract may also be achieved by means of an aerosol formulation in which the active ingredient is provided in a pressurized pack with a suitable propellant. If the compounds of the present invention or pharmaceutical compositions comprising them are administered as aerosols (e.g., nasal aerosols, by inhalation), this can be carried out, for example, using a spray, a nebulizer, a pump nebulizer, an inhalation apparatus, a metered inhaler or a dry powder inhaler. Pharmaceutical forms for administration of the compounds of the present invention as an aerosol can be prepared by processes well known to the person skilled in the art. Solutions or dispersions of the compounds of the present invention or a pharmaceutically acceptable salt, solvate, hydrate or derivative thereof in water, water/alcohol mixtures or suitable saline solutions, for example, can be employed using customary additives (e.g., benzyl alcohol or other suitable preservatives), absorption enhancers for increasing the bioavailability, solubilizers, dispersants and others and, if appropriate, customary propellants (e.g., carbon dioxide, CFCs, such as, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and the like). The aerosol may conveniently also contain a surfactant such as lecithin. The dose of drug may be controlled by provision of a metered valve.

In formulations intended for administration to the respiratory tract, including intranasal formulations, the compound will generally have a small particle size for example of the order of 10 microns or less. Such a particle size may be obtained by means known in the art, for example by micronization. When desired, formulations adapted to give sustained release of the active ingredient may be employed.

Alternatively, the active ingredients may be provided in the form of a dry powder (e.g., a powder mix of the compound in a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP)) Conveniently the powder carrier will form a gel in the nasal cavity. The powder composition may be presented in unit dose form (e.g., capsules, cartridges) as for gelatin or blister packs from which the powder may be administered by means of an inhaler.

The pharmaceutical preparations are preferably in unit dosage forms. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

In some embodiments, the compositions are tablets or capsules for oral administration.

In some embodiments, the compositions are liquids for intravenous administration.

The compounds according to the invention may optionally exist as pharmaceutically acceptable salts including pharmaceutically acceptable acid addition salts prepared from pharmaceutically acceptable non-toxic acids including inorganic and organic acids. Representative acids include, but are not limited to, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethenesulfonic, dichloroacetic, formic, fumaric, gluconic, glutamic, hippuric, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, oxalic, pamoic, pantothenic, phosphoric, succinic, sulfiric, tartaric, oxalic, p-toluenesulfonic and the like, such as those pharmaceutically acceptable salts listed by Berge et al., Journal of Pharmaceutical Sciences, 66:1-19 (1977), incorporated herein by reference in its entirety.

The acid addition salts may be obtained as the direct products of compound synthesis. In the alternative, the free base may be dissolved in a suitable solvent containing the appropriate acid and the salt isolated by evaporating the solvent or otherwise separating the salt and solvent. The compounds of this invention may form solvates with standard low molecular weight solvents using methods known to the skilled artisan.

Compounds of the present invention can be converted to “pro-drugs.” The term “pro-drugs” refers to compounds that have been modified with specific chemical groups known in the art and that when administered into an individual undergo biotransformation to give the parent compound. Pro-drugs can thus be viewed as compounds of the invention containing one or more specialized non-toxic protective groups used in a transient manner to alter or to eliminate a property of the compound. In one general aspect, the “pro-drug” approach is utilized to facilitate oral absorption. A thorough discussion is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems Vol. 14 of the A.C.S. Symposium Series; and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are hereby incorporated by reference in their entirety.

Some embodiments of the present invention include a method of producing a pharmaceutical composition for “combination-therapy” comprising admixing at least one compound according to any of the compound embodiments disclosed herein, together with at least one known pharmaceutical agent as described herein and a pharmaceutically acceptable carrier.

As will be recognized, the steps of the methods of the present invention need not be performed any particular number of times or in any particular sequence. Additional objects, advantages and novel features of this invention will become apparent to those skilled in the art upon examination of the following examples thereof, which are intended to be illustrative and not intended to be limiting.

EXAMPLES Example 1: Preparation of Compounds

The preparation of Compound 1, including certain crystal forms of Compound 1 are described in International Patent Application No. PCT/US2009/004265, published as International Publication No. WO2010/011316; and International Patent Application No. PCT/US2011/000153, published as International Publication No. WO2011/094008; the entire contents of each are incorporated herein by reference in their entirety.

The preparation of the crystal form and crystalline free-plate habit of the non-solvated L-arginine salt of Compound 1 is described in International Patent Application No. PCT/US2016/038506, published as International Publication No. WO2016/209809, the entire contents of which are incorporated herein by reference in their entirety.

Example 2 Clinical Trial for Treating Primary Biliary Cholangitis (PBC) With Compound 1

A phase I/II, open-label, single arm, proof-of-concept study is performed to evaluate the safety, tolerability and efficacy of Compound 1 in individuals with primary biliary cholangitis (PBC). The study is conducted in individuals aged 18 to 80 years old (inclusive) who have PBC and an inadequate response to ursodeoxycholic acid (UDCA) and are on a stable dose of UDCA for at least 6 months prior to screening. The trial includes an initial pilot study to assess pharmacokinetics and tolerability for up to ten individuals.

Primary objectives include efficacy (i.e., changes in ALP levels from baseline), safety and tolerability of Compound 1 in individuals with PBC over a 24-week treatment period.

Exploratory objectives include assessment of the following: 1) the pharmacokinetic (PK) profile of Compound 1 in individuals with PBC; 2) the pharmacodynamic (PD) response (lymphocyte counts and subsets thereof) and changes in antinuclear antibodies (ANA) (anti-gp210 and anti-sp100) and other exploratory biomarkers over a 24-week treatment period; 3) serum high sensitivity C-reactive protein (hsCRP), alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (GGT), antimitochondrial antibodies (AMA), Golgi protein 73 (GP73), complement factor 4 (C4), bile acid, total IgG, total IgM, over a 24-week treatment period; 4) complete blood count (CBC); 5) quality of life (QoL) and incidence of pruritus and/or fatigue over a 24-week treatment period; 6) safety, tolerability and efficacy of Compound 1 over a 12-week period; 7) liver stiffness using transient elastography over a 24-week treatment period; and 8) changes in eye dryness, the Schirmer test and tear breakup time (TBUT) test over a 24-week treatment period in individuals with abnormal results at screening.

Individuals are screened for up to four weeks, then administered 1 or 2 mg of Compound 1 for 24 consecutive weeks. Safety laboratory parameters, vital signs, physical examination, and concomitant medications are evaluated during the screening period, and Child-Pugh scores may be calculated. The trial is conducted as follows:

1) A four-week pilot study is run to assess safety and tolerability, including PK and maximum tolerated dose, in up to 10 individuals before proceeding with the remainder of the clinical trial. Individuals are admitted to the clinic on Day -1 and stay until Day 3. Vital signs and EKG changes are monitored extensively during this time. Individuals have abbreviated PK sampling pre-dose (baseline) and at 0.5, 1, 2, 4, 6, 8, and 12 hours after dosing on Day 1, and 6 hours post-dosing on Days 2 and 3. Individuals receive 1 mg of Compound 1 (PO, QD) on Days 1, 2, and 3. If 1 mg of Compound 1 is well tolerated and there are no clinically significant changes in vital signs and EKG, the 1 mg dose level is maintained and the individual is discharged on Day 3. If 1 mg is not tolerated, then treatment is stopped if any of the following findings are made during treatment:

-   Certain ECG changes (e.g., if a QTc interval ≥ 500 msec is observed     and confirmed, extended monitoring is performed until the findings     have resolved and dosing is stopped); -   ALT/AST >3 x baseline level and >ULN; or -   Total bilirubin >2 x baseline level and >1.5 mg/dL.

The 1 mg dose level may be increased to 2 mg at Week 4 based on the individual’s safety and PK data. Individuals with dose escalation on Week 4 have an additional PK sampling, and vital signs and ECG monitoring are performed every 30 minutes until at least 6 hours post dosing. Dosage is maintained at 2 mg QD if possible, but may return to 1 mg QD if deemed necessary.

2) Based on the above pilot study data, subsequent individuals are admitted to the clinic on Day -1 and start treatment on Day 1 (1 mg QD). PK sampling is performed up to 45 minutes pre-dose, and at 0.5, 1, 2, 4, 6, and 8 hours post dosing on Day 1. Vital signs and EKG changes are closely monitored. Additional PK samples are collected pre-dose on Weeks 1, 2, 4, 8, 12, 16, 20, and 24. The 1 mg dose level may be increased to 2 mg at Week 4 based on the individual’s safety and PK data. If 1 mg is not tolerated, treatment is stopped as described above. Individuals with dose escalation on Week 4 have an additional PK sampling, and vital signs and ECG monitoring are performed every 30 minutes until at least 6 hours post dosing. Dosage is maintained at 2 mg QD if possible, but may return to 1 mg QD if deemed necessary. Safety and tolerability are assessed by monitoring adverse events and vital signs, ECG, and blood tests. Individuals return to the study site at Weeks 1, 2, 4, 8, 12, 16, 20, and 24 for examinations as described in Schedule of Procedures and Visits (FIGS. 2 to 6 ).

The last dose of Compound 1 is administered one day before the end of the 24-week treatment period.

Individuals return to the study site for the final visit two weeks after the end of 24-week treatment, and final procedures are performed per Schedule of Procedures and Visits (FIGS. 2 to 6 ).

ALP, GGT, ALT/AST, bilirubin (total and direct), PT/INR, albumin, lipid panel, and total serum IgG/IgM as well as a PBC-40 questionnaire and a 5-D Pruritus scale/VAS are measured during the study period.

Individuals must meet the inclusion and exclusion criteria described below to be enrolled in the study.

Inclusion Criteria:

-   1. Individuals must be males or females aged 18 to 80 years     (inclusive) at the time of screening, with confirmed PBC diagnosis     based upon at least two of the following three criteria:     -   anti-mitochondrial antibody (AMA) titer > 1:40;     -   alkaline phosphate (ALP) > 1.5 x ULN for at least 6 months; and     -   liver biopsy findings consistent with PBC. -   2. An inadequate response to UDCA, as defined by an alkaline     phosphate (ALP) > 1.67 x ULN after 6 months of UDCA at a minimum     dose of 13 mg/kg/day. -   3. Individuals must be on a stable dose of UDCA for at least 3     months prior to screening. -   4. Individuals taking medications for pruritus or fatigue must have     been on stable doses of these medications for at least two weeks     prior to Day 1. -   5. Individuals must have the following laboratory parameters at     screening:     -   ALP > 1.67 x ULN but < 10 x ULN     -   ALT and AST <5 x ULN     -   total bilirubin < ULN     -   international normalized ratio (INR) < 1.2 x ULN     -   platelet count > 150,000 /mm³ (> 150 x 10⁹/L)     -   serum albumin >3.0 g/dL ( > 30 g/L)     -   serum creatinine < 1.5 mg/dL (133 µmol/L)     -   estimated glomerular filtration rate (eGFR) ≥ 60 mL/min/1.73 m²     -   TSH < 5.0 mU/L without clinical significant changes of free T3         and T4 levels     -   Fibroscan® (transient elastography) < 10 kPa -   6. Individuals are considered to be in stable health in the opinion     of the investigator as determined by:     -   a) a screening physical examination with no clinically         significant abnormalities unrelated to PBC.     -   b) vital signs at screening: pulse rate ≥ 55 bpm, systolic blood         pressure (SBP) ≥ 90, and diastolic blood pressure (DBP) ≥ 55         mmHg.     -   c) no clinical abnormalities noted in the 12-lead         electrocardiogram (ECG) in the opinion of the investigator (see         also exclusion criteria #22 and #23).     -   d) no evidence of macular edema in an ophthalmology evaluation         (performed by an ophthalmologist), supported with optical         coherence tomography (OCT), where available (dependent on site         capability) at screening or no later than three months prior to         screening.

Exclusion Criteria:

-   1. Chronic liver disease of a non-PBC etiology. However, PBC     patients accompanied with primary Sjögren’s syndrome (pSS) are     eligible to be enrolled. -   2. History or evidence of clinically significant hepatic     decompensation:     -   a) Portal hypertension, cirrhosis and complications of         cirrhosis/portal hypertension (e.g., variceal hemorrhage,         encephalopathy or ascites).     -   b) History of liver transplantation, current placement on a         liver transplant list or current Model for End Stage Liver         Disease (MELD) score > 12.     -   c) Cirrhosis with complications, including history or presence         of: spontaneous bacterial peritonitis, hepatocellular carcinoma,         hyperbilirubinemia > 1.5 x ULN.     -   d) Hepatorenal syndrome (type I or II).     -   e) Splenomegaly. -   3. Medical conditions that may cause non-hepatic increases in ALP     (e.g., Paget’s disease). -   4. No evidence of worsening liver function during the screening     period. -   5. Patients who have donated any blood, or had significant blood     loss within 30 days prior to screening. -   6. Clinically significant infections (e.g., pneumonia,     pyelonephritis) as judged by the investigator with an end date less     than 6-weeks prior to treatment start (Day 1). In case of infection     requiring hospitalization or intravenous antimicrobial therapy, or     opportunistic infections, those infections must have ended at least     8 weeks prior to Day 1. -   7. Infection with hepatitis C virus anytime in the past; confirmed     active infection with hepatitis B virus at screening. -   8. Current active or latent tuberculosis (TB) or history of TB that     has not been successfully treated. In case of documented successful     treatment in the past and current positivity of the QuantiFERON®     test this criterion may be assessed on a case by case basis with     medical monitor. -   9. A positive diagnostic TB test at screening defined as a positive     QuantiFERON® test or 2 successive indeterminate QuantiFERON® tests. -   10. Agents used for the treatment of any condition listed in the     exclusionary enrollment criteria from 30 days prior to Day 1. -   11. Exposure to B cell or T cell targeted therapies (such as     natalizumab, rituximab, abatacept, ustekinumab) within 30 days prior     to Day 1. -   12. Azathioprine, colchicine, or methotrexate within 30 days prior     to Day 1. -   13. Treatment with obeticholic acid (OCA) or fibrates (including     bezafibrate) within 30 days prior to Day 1. -   14. Other immunosuppressive, immunomodulating or antineoplastic     agents within 30 days prior to Day 1 (or not meeting the stability     time period for concomitant medications indicated as permitted).     However, medications (such as for pruritus and fatigue) used as     adjunctive therapy for PBC in combination with UDCA should be at     stable doses within 30 days prior to Day 1. These medications may be     adjusted during study treatment, but all adjustments must be     recorded in the concomitant medications source documents and eCRFs.

Drugs not allowed per the UDCA prescribing information: agents used for the treatment of any condition listed in the exclusionary enrollment criteria within 30 days prior to Day 1; Exposure to B cell or T cell targeted therapies (such as natalizumab, rituximab, abatacept, ustekinumab) within 30 days prior to Day 1 and during the study; azathioprine, colchicine, or methotrexate within 30 days prior to Day 1 and during the study; fibrates (including bezafibrate) within 30 days prior to Day 1 and during the study; treatment with OCA within 30 days prior to Day 1 and during the study; immunosuppressive, immunomodulating agents such as 5-ASA, azathioprine, colchicine, or methotrexate due to the potential impact on ALP; use of moderate to strong inhibitors of CYP2C9 (e.g., amiodarone, felbamate, fluconazole, miconazole, piperine, diosmin, disulfiram..m, fluvastatin, fluvoxamine, voriconazole) within 30 days prior to Day 1; receipt of live vaccine from 30 days prior to Day 1 (and for at least 6 months after the last dose of study drug); receipt of any investigational agent (including S1P modulators) within 30 days or 5 half-lives (whichever is longer), prior to Day 1; investigational agents, other than Compound 1 from 30 days prior to Day 1 and during the study; and receipt of any other medications that in the opinion of the investigator precludes patient from the safe participation in the study.

15. Use of moderate to strong inhibitors of CYP2C9 (e.g., amiodarone, felbamate, fluconazole, miconazole, piperine, diosmin, disulfiram, fluvastatin, fluvoxamine, voriconazole) within 30 days prior to Day 1.

16. Patients without documented positive varicella zoster virus (VZV) IgG-antibody status or patients who have not completed VZV vaccination within 6 weeks prior to Day 1.

17. Receipt of live vaccine within 30 days prior to Day 1 (and for at least 6 months after the last dose of study drug).

18. Receipt of any investigational agent (including S1P modulators) within 30 days or 5 half-lives (whichever is longer), prior to Day 1.

19. Receipt of any other medication that in the opinion of the investigator precludes patients from the safe participation in the study.

20. Abnormal forced expiratory volume (FEV₁) or forced vital capacity (FVC), i.e., < 80% of predicted values at screening.

21. Any known history of congenital or acquired immuno- deficiency (e.g., common variable immunodeficiency, human immunodeficiency virus [HIV] infection [ELISA and Western blot] test result, organ transplantation).

22. Recent history (within 6 months of screening visit) of cardio- or cerebrovascular disease, acute coronary syndrome (ACS), myocardial infarction (MI), cardiomyopathy, heart failure, unstable angina, cerebro-vascular accident, including transient ischemic attack (TIA).

23. History or presence of cardiac arrhythmia, conduction system disease (including AV node dysfunction, 2nd or 3rd degree heart block, and sick sinus syndrome), or use of Class Ia or Class III anti arrhythmic agents, or baseline QTc >500 msec.

24. Any surgical procedure requiring general anesthesia within 30 days prior to Day 1 or plans to undergo major surgery during the study period.

25. History or presence of retinal macular edema.

26. History of or signs and symptoms of progressive multifocal leukoencephalopathy (PML) as assessed by the PML checklist at screening. If PML is suspected, withhold dosing and refer to a neurologist; if confirmed, discontinue dosing permanently.

27. History of more than one episode of herpes zoster or any episode of disseminated zoster.

28. History of lymphoproliferative disorder, lymphoma, leukemia, myeloproliferative disorder, or multiple myeloma.

29. Leukopenia or lymphopenia at screening.

30. History of malignancy except for adequately treated basal cell skin cancer and in situ carcinoma of the cervix or of the uterus that have been completely excised with documented, clear margins.

31. History of severe allergic or anaphylactic reactions requiring medical attention.

32. History of uncontrolled hypothyroidism.

33. Current or recent history (within 1 year prior to Day 1) of alcohol dependence or illicit drug use.

34. Active psychiatric disorders that, in the investigator’s opinion, may interfere with compliance, with the study procedures.

35. History of any other clinically significant medical condition that, in the investigator’s opinion, would preclude patients from safe participation in the study.

36. Inability to attend all the study visits or comply with study procedures.

Example 3: BET (Brunauer, Emmett, and Teller) Specific Surface Area Method (Plate Habit)

In general, the specific surface areas for crystalline free-plate habit of the non-solvated L-arginine salt of Compound 1 were determined by physical adsorption of nitrogen gas on the surface of the sample from each lot using the well-established technique based on the Brunauer, Emmett, and Teller theory.

The BET surface areas for the samples were measured by Micromeritics Pharmaceutical Services using a Micromeritics™ TriStar II BET surface area analyzer (MicroActive for TriStar II Plus 2.02 Software™). The samples were degassed at 25° C. for 960 minutes under vacuum (i.e., 100 mm/Hg). The determination of the adsorption of N₂ at 77.3 K was measured using a BET surface area eleven-point method with relative pressures in the range of about 0.05 to about 0.3 (P/P₀) for a weighed amount of each sample, see Table 1 below. The analysis was performed per ISO9277.

TABLE 1 Arena Lot Number Lot Number Sample (g) Correlation Coefficient BET Surface Area (m²/g) Isolated Morphology DSC Onset Temperature 5015-12-12 A1 0.6163 0.99916 0.7 Plates 208.09° C. 5015-12-13 A2 1.5270 0.99945 0.7 Plates 207.20° C. 5015-12-14 A3 0.4465 0.99922 1.5 Plates 207.19° C. 5015-12-15 A4 0.5709 0.99939 1.0 Plates 207.83° C. 5015-12-16 A5 0.9582 0.99940 0.8 Plates 207.90° C. 04GSp A6 0.4332 0.99921 2.4 Plates 206.55° C. 05GSp A7 0.3652 0.9991 1.9 Plates 206.94° C. 06GSp A8 0.6866 0.99984 3.0 Plates 207.04° C. 07GSp A9 0.2754 0.99914 3.1 Plates 207.63° C.

Example 4 Formulations for L-Arginine Salt of (R)-2-(7-(4-Cyclopentyl-3-(trifluoromethyl)-Benzyloxy)-1,2,3,4-Tetrahydrocyclopenta[b]indol-3-yl)Acetic Acid

Core tablets were manufactured using the formulation as described in Table 2 and using substantially the same process as described in FIG. 7 . The L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is 72.42% free acid (Compound 1) and 27.58% L-arginine (i.e., 1.381 mg of the L-arginine salt of Compound 1 corresponds to 1 mg of active/free acid).

TABLE 2 Tablet Strength 1 mg 2 mg L-Arg Salt of Compound 1 1.381 2.762 Mannitol Pearlitol® 100 SD 54.119 52.738 Microcrystalline cellulose - Avicel® 40 40 Sodium Starch Glycolate - Explotab® 4 4 Magnesium Stearate 0.5 0.5 Opadry® II Blue 4 4 Total tablet target weight 104 104

Those skilled in the art will recognize that various modifications, additions, substitutions, and variations to the illustrative examples set forth herein can be made without departing from the spirit of the invention and are, therefore, considered within the scope of the invention. 

1. A method of treating primary biliary cholangitis (PBC) in an individual in need thereof comprising administering a therapeutically effective amount of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1), or a pharmaceutically salt, solvate, or hydrate thereof, the method comprising: (a) analyzing one or more samples from the individual for a first level of at least one biomarker obtained prior to the treatment with Compound 1; (b) administering Compound 1 to the individual; (c) analyzing one or more samples from the individual for a second level of the at least one biomarker obtained after the treatment with Compound 1; and (d) (i) continuing administration of Compound 1 if the second level of the at least one biomarker in step (c) is less than or about equal to the corresponding first level of the at least one biomarker in step (a); or (ii) discontinuing administration of Compound 1 if the second level of the at least one biomarker in step (c) is greater than the corresponding first level of the at least one biomarker in step (a); wherein the at least one biomarker is selected from the group consisting of: (i) anti-gp210; (ii) anti-sp100; (iii) serum high sensitivity C-reactive protein (hsCRP), (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM.
 2. (canceled)
 3. The method according to claim 1, wherein the individual was previously treated with a therapeutically effective amount of ursodeoxycholic acid (UDCA).
 4. The method according to claim 1, wherein the individual is currently treated with a therapeutically effective amount of ursodeoxycholic acid (UDCA).
 5. (canceled)
 6. The method according to claim 1, wherein the individual was previously treated with ursodeoxycholic acid (UDCA) and the individual had an inadequate response to UDCA.
 7. The method or use according to claim 6, wherein the individual had an inadequate response to UDCA as determined by an alkaline phosphate (ALP)>1.67x upper limit of normal (ULN) for the individual. 8-14. (canceled)
 15. The method according to claim 1, wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is administered orally.
 16. (canceled)
 17. The method according to claim 1, wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1.0 mg to about 5 mg of Compound
 1. 18. The method according to claim 1, wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg to about 2 mg.
 19. The method according to claim 1 wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, or about 2 mg of Compound
 1. 20. The method according to claim 1 , wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 1 mg of Compound
 1. 21. The method according to claim 1 , wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is in an amount equivalent to about 2 mg of Compound
 1. 22. The method according to claim 1 , wherein the therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is administered once daily.
 23. The method according to claim 1 , wherein the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an L-arginine salt of Compound
 1. 24. The method according to claim 1 , wherein the Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, is an anhydrous, non-solvated crystalline form of the L-arginine salt of Compound
 1. 25-31. (canceled)
 32. A method of treating an individual in need thereof with (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclo-penta[b]indol-3-yl)acetic acid (Compound 1) comprising: (a) administering Compound 1 to the individual; (b) analyzing one or more samples from the individual for the level of at least one biomarker obtained after the treatment with Compound 1 ; and (c) (i) continuing administration of Compound 1 if the at least one biomarker is less than or equal to a predetermined level for the at least one biomarker prior to treatment of Compound 1 ; or (ii) discontinuing administration of Compound 1 if the at least one biomarker is greater than a predetermined level for the at least one biomarker prior to treatment of Compound 1 ; wherein the at least one biomarker is selected from the group consisting of: (i) anti-gp210; (ii) anti-splOO; (iii) serum high sensitivity C-reactive protein (hsCRP), (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM. 33-40. (canceled)
 41. The method of claim 1, wherein the second level of the at least one biomarker in step (c) is 2% less, 5% less, 10% less, 12% less, 15% less, 17% less, 20% less, 22% less, 25% less, 30% less, 35% less, 40% less, 45% less, 50% less, or > 50% less than the corresponding first level of the at least one biomarker in step (a).
 42. The method of claim 1, wherein the at least one biomarker is two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve biomarkers selected from the group consisting of: (i) anti-gp210; (ii) anti-splOO; (iii) serum high sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM.
 43. The method of claim 1, wherein the at least one biomarker in step (c) is two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve biomarkers selected from the group consisting of: (i) anti-gp210; (ii) anti-splOO; (iii) serum high sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM.
 44. The method of claim 32, wherein the individual has primary biliary cholangitis (PBC).
 45. The method of claim 32, wherein the individual has fatigue, pruritus, eye dryness, and/or Sjogren’s syndrome (SS).
 46. The method of claim 32, wherein the at least one biomarker is two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve biomarkers selected from the group consisting of: (i) anti-gp210; (ii) anti-splOO; (iii) serum high sensitivity C-reactive protein (hsCRP); (iv) alanine transaminase (ALT); (v) aspartate transaminase (AST); (vi) gamma-glutamyl transferase (GGT); (vii) antimitochondrial antibodies (AMA); (viii) Golgi protein 73 (GP73); (viii) bile acid; (x) complement factor 4 (C4); (xi) IgG; and (xii) IgM. 